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<p>Multiplex polymerase chain reaction (PCR) tests multiple biomarkers or pathogens that cause overlapping symptoms, making it an essential tool in syndromic testing. To achieve a multiplex PCR on chip, a design based on capillary-driven fluidic actuation is proposed. Our silicon chip features 22 reaction chambers and allows primers and probes to be pre-spotted in the reaction chambers prior to use. The design facilitates rapid sample loading through a common inlet channel, delivering reagents to all reaction chambers in less than 10&#xa0;s. A custom clamping mechanism combined with a double depth cavity design ensures proper sealing during temperature cycling without the need for extra reagents like oil. Temperature cycling and fluorescence imaging were performed using custom-made hardware. As a proof of concept, two single nucleotide polymorphisms (SNPs), CyP2C19*2 and PCSK9 were detected. These results demonstrate the feasibility of on-chip multiplex PCR, compatible with different assays in parallel and requiring only a single pipetting step for reagent loading, without active fluidic actuation like pumping.</p>

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A capillary-driven microfluidic device for performing spatial multiplex PCR

  • 期刊论文

  • 发表日期：

    2025年3月26日

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  • Rodrigo S. Wiederkehr,
  • Elisabeth Marchal,
  • Maarten Fauvart,
  • Tomas Forceville,
  • Ahmed Taher,
  • Tim Steylaerts,
  • YoungJae Choe,
  • Hans Dusar,
  • Silvia Lenci,
  • Eleni Siouti,
  • Vassiliki T. Potsika,
  • Evangelos Andreakos,
  • Tim Stakenborg

摘要

Multiplex polymerase chain reaction (PCR) tests multiple biomarkers or pathogens that cause overlapping symptoms, making it an essential tool in syndromic testing. To achieve a multiplex PCR on chip, a design based on capillary-driven fluidic actuation is proposed. Our silicon chip features 22 reaction chambers and allows primers and probes to be pre-spotted in the reaction chambers prior to use. The design facilitates rapid sample loading through a common inlet channel, delivering reagents to all reaction chambers in less than 10 s. A custom clamping mechanism combined with a double depth cavity design ensures proper sealing during temperature cycling without the need for extra reagents like oil. Temperature cycling and fluorescence imaging were performed using custom-made hardware. As a proof of concept, two single nucleotide polymorphisms (SNPs), CyP2C19*2 and PCSK9 were detected. These results demonstrate the feasibility of on-chip multiplex PCR, compatible with different assays in parallel and requiring only a single pipetting step for reagent loading, without active fluidic actuation like pumping.